This proposal seeks to develop a novel, highly miniaturized peptide microarray for the simultaneous profiling of multiple protein kinases in a high throughput, multiplexed format. A universal, nonprotein, dye conjugation method of identifying phosphorylated peptide substrates will be developed that will provide for a direct correlation of phosphate incorporation as a result of kinase activity. This platform is unique since it will allow for the screening of small molecules that will inhibit only certain kinases while in the presence of multiple other kinases due to the multiplex format of the assay. This will not only increase the amount of information gained per assay based on today's standards, it will also provide for testing the specificity of the potential inhibitor due to the presence of multiple kinases in the same assay sample. This technology can be applied to many scientific areas involved in the study of kinases including cellular signal transduction and high throughput drug screening of small molecule inhibitors. Other basic and diagnostic applications of this technology will enable the careful examination of the molecular basis for many human diseases and chronic conditions including Alzheimer's disease, cancer, and diabetes. [unreadable] [unreadable] [unreadable]